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Sci Rep ; 7(1): 3142, 2017 06 09.
Artigo em Inglês | MEDLINE | ID: mdl-28600512

RESUMO

Intestinal bacterial ß-glucuronidase (ßG) hydrolyzes glucuronidated metabolites to their toxic form in intestines, resulting in intestinal damage. The development of a method to inhibit ßG is thus important but has been limited by the difficulty of directly assessing enzyme activity in live animals. Here, we utilized a fluorescent probe, fluorescein di-ß-D-glucuronide (FDGlcU), to non-invasively image the intestinal bacterial ßG activity in nude mice. In vitro cell-based assays showed that the detection limit is 104 colony-forming units/well of ßG-expressing bacteria, and that 7.81 ng/mL of FDGlcU is enough to generate significant fluorescent signal. In whole-body optical images of nude mice, the maximum fluorescence signal for ßG activity in intestines was detected 3 hours after gavage with FDGlcU. Following pretreatment with a bacterial ßG inhibitor, the fluorescence signal was significantly reduced in abdomens and excised intestines images. For a 4-day antibiotic treatment to deplete intestinal bacteria, the FDGlcU-based images showed that the ßG activity was decreased by 8.5-fold on day 4 and then gradually increased after treatment stopped. The results suggested that FDGlcU-based imaging revealed the in vitro and in vivo activity of intestinal bacterial ßG, which would facilitate pharmacodynamic studies of specific bacterial ßG inhibitors in animal studies.


Assuntos
Bactérias/enzimologia , Fluoresceínas/química , Corantes Fluorescentes/química , Glucuronidase/metabolismo , Intestinos/microbiologia , Imagem com Lapso de Tempo/métodos , Animais , Antibacterianos/administração & dosagem , Antibacterianos/farmacocinética , Bactérias/efeitos dos fármacos , Proteínas de Bactérias/metabolismo , Fluoresceínas/administração & dosagem , Corantes Fluorescentes/administração & dosagem , Hidrólise , Intestinos/efeitos dos fármacos , Limite de Detecção , Camundongos , Camundongos Nus , Imagem Molecular
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